Abstract
Cyclosporin A (CyA) has received meticulous attention owing to its immunosuppressive and biological activities. In this study, a soil isolate, capable of producing CyA, was named Zag1 strain and identified as Aspergillus fumigatus based on macroscopic and microscopic characteristics, 18S rDNA sequence, and phylogenetic characteristic analysis. To maximize the production of CyA, the fungal culture was grown under various fermentation conditions including selection of the cultivation medium, agitation rate, fermentation time, incubation temperature, pH value, inoculum nature, and medium volume. A simple medium (pH 5.0) containing 5% maltose as a carbon source and 2% potassium nitrate as a nitrogen source favored the highest CyA production when the fermentation process was maintained at 120 rpm for 9 days and at 30 °C using 3% standard inoculum of 5-day-old. The final CyA titer under these conditions was intensified to 2.23–3.31-fold, as compared with the amount obtained with seven types of basal media. A. fumigatus Zag1 appears to possess a good biotechnological potential for CyA production under favorable culture conditions.
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References
Aarnio TH, Agathos SN (1989) Production of extracellular enzymes and cyclosporin by Tolypocladium inflatum and morphologically related fungi. Biotechnol Lett 11:759–764
Abdel-fattah YR, El Enshasy H, Anwar M, Omar H, Abolmagd E, Abou Zahra R (2007) Application of factorial experimental designs for optimization of cyclosporin A production by Tolypocladium inflatum in submerged culture. J Microbiol Biotechnol 17:1930–1936
Agathos SN, Parekh R (1990) Enhancement of cyclosporin production in a Tolypocladium inflatum strain after epichlorohydrin treatment. J Biotechnol 13:73–81
Agathos SN, Marshall JW, Moraiti C, Parekh R, Madhosingh C (1986) Physiological and genetic factors for process development of cyclosporine fermentations. J Ind Microbiol 1:39–48
Agathos SN, Madhosingh C, Marshall JW, Lee J (1987) The fungal production of cyclosporine. Ann N Y Acad Sci 506:657–662
Altschul SF, Madden TL, Schäffer AA, Zhang J, Zhang Z, Miller W, Lipman DJ (1997) Gapped BLAST and PSI-BLAST: a new generation of protein database search programs. Nucleic Acids Res 25:3389–3402
Amor KT, Ryan C, Menter A (2010) The use of cyclosporine in dermatology: part I. J Am Acad Dermatol 63:925–946
Ando O, Satake H, Nakajima M, Sato A, Nakamura T, Kinoshita T, Furuya K, Haneishi T (1991) Synerazol, a new antifungal antibiotic. J Antibiot 44:382–389
Anjum T, Azam A, Irum W (2012) Production of cyclosporine A by submerged fermentation from a local isolate of Penicillium fellutanum. Indian J Pharm Sci 74:372–374
Balakrishnan K, Pandey A (1996a) Growth and cyclosporin A production by an indigenously isolated strain of Tolypocladium inflatum. Folia Microbiol 41:401–406
Balakrishnan K, Pandey A (1996b) Influence of amino acids on the biosynthesis of cyclosporin A by Tolypocladium inflatum. Appl Microbiol Biotechnol 45:800–803
Balaraman K, Mathew N (2006) Optimization of media composition for the production of cyclosporin A by Tolypocladium species. Indian J Med Res 123:525–530
Billich A, Zocher R (1987) Enzymatic synthesis of cyclosporin A. J Biol Chem 262:17258–17259
Debeaupuis JP, Sarfati J, Chazalet V, Latgé JP (1997) Genetic diversity among clinical and environmental isolates of Aspergillus fumigatus. Infect Immun 65:3080–3085
Dreyfuss MM, Gams W (1994) Proposal to reject Pachybasium niveum Rostrup in order to retain the name Tolypocladium inflatum for the fungus that produces cyclosporin. Taxon 34:660–661
Dreyfuss M, Härri E, Hofmann H, Kobel H, Pache W, Tscherter H (1976) Cyclosporin A and C: new metabolites from Trichoderma polysporum (Link ex Pers.) Rifai. Eur J Appl Microbiol Biotechnol 3:125–133
El-Refai NA, Abd-Elsalam IS, Sallam LA (2004) Kinetic studies on the growth and cyclosporin A production by a local isolate of Fusarium oxysporum NRC. Acta Pharm Turcica 46:197–204
Ferraccioli GF, Tomietto P, De Santis M (2005) Rationale for T cell inhibition by cyclosporine A in major autoimmune diseases. Ann N Y Acad Sci 1051:658–665
Furtado NA, Said S, Ito IY, Bastos JK (2002) The antimicrobial activity of Aspergillus fumigatus is enhanced by a pool of bacteria. Microbiol Res 157:207–211
Gams W (1971) Tolypocladium. Eine Hyphomycetengattung mit geschwollenen Phialiden. Persoonia 6:185–191
Germano V, Picchianti Diamanti A, Ferlito C, Podestà E, Salemi S, Migliore A, D’Amelio R, Laganà B (2011) Cyclosporine A in the long-term management of systemic lupus erythematosus. J Biol Regul Homeost Agents 25:397–403
Isaac CE, Jones A, Pickard MA (1990) Production of cyclosporins by Tolypocladium niveum strains. Antimicrob Agents Chemother 34:121–127
Ismaiel AA, El-Sayed EA, Mahmoud AA (2010) Some optimal culture conditions for production of cyclosporin A by Fusarium roseum. Braz J Microbiol 41:1112–1123
Jegorov A, Matha V, Weiser J (1990) Production of cyclosporins by entomopathogenic fungi. Microbios Lett 45:65–69
Jóhannsdóttir S, Jansook P, Stefánsson E, Loftsson T (2015) Development of a cyclodextrin-based aqueous cyclosporin A eye drop formulations. Int J Pharm 493:86–95
Kadlec Z, Šimek P, Heydová A, Jegorov A, Mat’ha V, Landa Z, Eyal J (1994) Chemotaxonomic discrimination among the fungal genera Tolypocladium, Beauveria and Paecilomyces. Biochem Syst Ecol 22:803–806
Kitahara K, Kawai S (2007) Cyclosporine and tacrolimus for the treatment of rheumatoid arthritis. Curr Opin Rheumatol 19:238–245
Kobel H, Traber R (1982) Directed biosynthesis of cyclosporins. Eur J Appl Microbiol Biotechnol 14:237–240
Kreuzig F (1984) High-speed liquid chromatography with conventional instruments for the determination of cyclosporin A, B, C and D in fermentation broth. J Chromatogr A 290:181–186
Kundu AK, Das D, Gupta TK (1973) Influence of culture and nutritional conditions on the production of amylase by submerged culture of Aspergillus oryzae. J Ferment Technol 4:142–150
Lee J, Agathos SN (1991) Dynamics of L-valine in relation to the production of cyclosporin A by Tolypocladium inflatum. Appl Microbiol Biotechnol 34:513–517
Lee MJ, Lee HN, Han K, Kim ES (2008) Spore inoculum optimization to maximize cyclosporin A production in Tolypocladium niveum. J Microbiol Biotechnol 18:913–917
Margaritis A, Chahal PS (1989) Development of a fructose based medium for biosynthesis of cyclosporin A by Beauveria nivea. Biotechnol Lett 11:765–768
Moubasher AH (1993) Soil fungi in Qatar and other Arab countries. The Centre for Scientific and Applied Research, Doha
Moussaïf M, Jacques P, Schaarwächter P, Budzikiewicz H, Thonart P (1997) Cyclosporin C is the main antifungal compound produced by Acremonium luzulae. Appl Environ Microbiol 63:1739–1743
Mukhopadhyay T, Roy K, Coutinho L, Rupp RH, Ganguli BN, Fehlhaber HW (1987) Fumifungin, a new antifungal antibiotic from Aspergillus fumigatus Fresenius 1863. J Antibiot 40:1050–1052
Nakajima H, Hamasaki T, Nishimura K, Kimura Y, Udagawa S, Sato S (1988) Isolation of 2-acetylamino-3-hydroxy-4-methyl-oct-6- enoic acid, a derivative of the ‘C9 amino acid’ residue of cyclosporins, produced by the fungus Neocosmospora vasinfecta E. F. Smith. Agric Biol Chem 52:1621–1623
Ramana Murthy MV, Mohan EVS, Sadhukhan AK (1999) Cyclosporin A production by Tolypocladium inflatum using solid state fermentation. Process Biochem 34:269–280
Raper KB, Fennell DI (1965) The genus Aspergillus. The Williams and Wilkins Company, Baltimore
Rifai MA (1969) A revision of the genus Trichoderma. Mycol Papers 116:1–56
Ruijter GJ, Visser J (1997) Carbon repression in Aspergilli. FEMS Microbiol Lett 151:103–114
Sallam LAR, El-Refai AH, Hamdy AA, El-Minofi AH, Abd-Elsalam IS (2003) Role of some fermentation parameters on cyclosporin A production by a new isolate of Aspergillus terreus. J Gen Appl Microbiol 49:321–328
Samson RA, Soares GG Jr (1984) Entomopathogenic species of the hyphomycete genus Tolypocladium. J Invertebr Pathol 43:133–139
Sawai K, Okuno T, Tereda Y, Harada Y, Sawamura K, Sasaki H, Takao S (1981) Isolation and properties of two antifungal substances from Fusarium solani. Agric Biol Chem 45:1223–1228
Serrano R, Gusmão L, Amorim A, Araujo R (2011) Rapid identification of Aspergillus fumigatus within the section Fumigati. BMC Microbiol 11:82. doi:10.1186/1471-2180-11-82
Son KH, Kim YK, Lee HW, Lee WS, Kim SU, Jeong TS, Kwon BM, Bok SH (1996) Amplification of farnesyl protein transferase inhibitory activity from Aspergillus fumigatus F93 by Plackett-Burman design. Biotechnol Lett 18:1297–1300
Survase SA, Kagliwal LD, Annapure US, Singhal RS (2011) Cyclosporin A—a review on fermentative production, downstream processing and pharmacological applications. Biotechnol Adv 29:418–435
Traber R, Hofmann H, Kobel H (1989) Cyclosporins—new analogues by precursor directed biosynthesis. J Antibiot 42:591–597
von Arx JA (1986) Tolypocladium, a synonym of Beauveria. Mycotaxon 25:153–158
Wenger RM, Payne TG, Schreier MH (1986) Cyclosporine: chemistry, structure-activity relationships and mode of action. Prog Clin Biochem Med 3:176–191
White TJ, Bruns T, Lee S, Taylor J (1990) Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis MA, Gelfand DH, Sinsky JJ, White TJ (eds) PCR protocols. A guide to methods and applications. Academic, San Diego
Zhao D, Beran M, Kozová J, Řeháček Z (1991) Formation of cyclosporins by Tolypocladium inflatum. Folia Microbiol 36:549–556
Zocher R, Nihira T, Paul E, Madry N, Peeters H, Kleinkauf H, Keller U (1986) Biosynthesis of cyclosporin A: partial purification and properties of a multifunctional enzyme from Tolypocladium inflatum. Biochemistry 25:550–553
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The author thanks Mr. El-Sayed R. El-Sayed, Plant Research Department, Nuclear Research Center, Atomic Energy Authority, Cairo, Egypt for performing the statistical analyses.
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Ismaiel, A.A. Production of the immunosuppressant cyclosporin A by a new soil isolate, Aspergillus fumigatus, in submerged culture. Appl Microbiol Biotechnol 101, 3305–3317 (2017). https://doi.org/10.1007/s00253-016-8052-0
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DOI: https://doi.org/10.1007/s00253-016-8052-0